Liquid-liquid phase separation and aggregation of the prion protein globular domain modulated by a high-affinity DNA aptamer

Matos, C. O.; Passos, Y. M.; Amaral, M. J.; Macedo, B.; Tempone, M.; Bezerra, O. C. L.; Moraes, M. O.; Almeida, M. S.; Weber, G.; Missailidis, S.; Silva, J. L.; Pinheiro, A. S.; Cordeiro, Y.

doi:10.1101/659037

bioRxiv · 10.1101/659037

Liquid-liquid phase separation and aggregation of the prion protein globular domain modulated by a high-affinity DNA aptamer

Matos, C. O.·Passos, Y. M.·Amaral, M. J.·Macedo, B.·Tempone, M.·Bezerra, O. C. L.·Moraes, M. O.·Almeida, M. S.·Weber, G.·Missailidis, S.·Silva, J. L.·Pinheiro, A. S.·Cordeiro, Y.

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Abstract

Structural conversion of cellular prion protein (PrPC) into scrapie PrP (PrPSc) and subsequent aggregation are key events for the onset of Transmissible Spongiform Encephalopathies (TSEs). Experimental evidences support the role of nucleic acids (NAs) in assisting the protein conversion process. Here, we used the SELEX methodology to identify two 25-mer DNA aptamers against the globular domain of recombinant murine PrP (rPrP90-231), namely A1 and A2. High-affinity binding of A1 and A2 to rPrP was verified by ITC. Aptamers structure was characterized by theoretical predictions, CD, NMR and SAXS, revealing that A1 adopts a hairpin conformation. Aptamer binding caused dynamic aggregation of rPrP90-231, resulting from the ability of rPrP90-231 to undergo liquid-liquid phase separation (LLPS). While free rPrP90-231 phase separated into large droplets, aptamer binding increased the amount but reduced the size of the condensates. Strikingly, a modified A1 aptamer that does not adopt a hairpin structure induced transition to an ordered state, suggestive of amyloid formation on the surface of the droplets. Our results describe for the first time PrP:NA interaction leading to LLPS and modulation of this effect depending on NA structure and binding stoichiometry, shedding light on the role of NAs in PrP misfolding and TSEs.

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Matos, C. O., Passos, Y. M., Amaral, M. J., Macedo, B., Tempone, M., Bezerra, O. C. L., Moraes, M. O., Almeida, M. S., Weber, G., Missailidis, S., Silva, J. L., Pinheiro, A. S., Cordeiro, Y.. 2019-06-03. Liquid-liquid phase separation and aggregation of the prion protein globular domain modulated by a high-affinity DNA aptamer. https://doi.org/10.1101/659037

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